Interaction between the product of the breast cancer susceptibility gene BRCA2 and DSS1, a protein functionally conserved from yeast to mammals.

Abstract

Coimmunoprecipitation of BRCA2 and DSS1 in mammalian cells. Marston N J et al. Mol. Cell. Biol. 1999;19:4633-4642 Coimmunoprecipitation of BRCA2 and DSS1 in mammalian cells. (A) DSS1 antisera can be used for specific immunoprecipitation. Preparations of GST or GST-DSS1 were immunoprecipitated in parallel with 1 or 10 μl of polyclonal antisera generated in this study. Antisera 286 and 287 were raised against the full-length recombinant DSS1, and antisera 290 and 291 were raised against a C-terminal peptide of DSS1. Immunoprecipitates were analyzed by immunoblotting with anti-GST antibody. The input proteins are shown. Sizes in both panels are indicated in kilodaltons. (B) Interaction of DSS1 with the C terminus of BRCA2 in transiently transfected COS cells. Lysates from COS cells transiently transfected with 9E10-tagged DSS1 (+) and various FLAG-tagged BRCA2 fragments (indicated by amino acid numbers) were immunoprecipitated with anti-Myc tag antibody 9E10, anti-DSS1 antiserum 290, or control anti-HA tag antibody 12CA5. Immunoprecipitates were analyzed by SDS-PAGE (6% polyacrylamide gel), together with samples of the input lysates representing half the protein used in the immunoprecipitations, followed by Western blot analysis with anti-FLAG tag antibody M2.

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